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Powdered Media Preparation Instructions

Preparation of Media for Filtration

Powdered media and salt mixtures are extremely hygroscopic and should be protected from atmospheric moisture. The entire contents of each package should be used immediately after opening. Preparing medium in concentrated form is not recommended as precipitates may form. Supplements can be added prior to filtration or introduced aseptically to sterile medium. The nature of the supplement may affect the storage conditions and shelf life of the medium.

Procedure

  • Measure out 90% of final volume of tissue culture grade water. Water should be at room temperature.
  • While gently stirring the water, add the powdered medium or salt mixture. Stir until dissolved. Do not heat water.
  • Rinse original package with a small amount of water to remove all traces of powder. Add to solution.
  • To the solution, add the required amount of sodium bicarbonate from the chart below for each liter of final volume of medium being prepared. Stir until dissolved.
  • While stirring, adjust the pH to 0.1-0.3 pH units below the desired pH since the pH may rise during vacuum filtration. The use of 1 N HCl or 1 N NaOH is recommended.
  • Bring medium to final volume with tissue culture grade water.
  • Sterilize immediately by filtration using a membrane with a porosity of 0.2 micrometers or less.
  • Aseptically dispense into sterile containers. Store liquid medium refrigerated at 2-8 °C and in the dark.
  • Osmolality will vary with the addition of sodium or potassium chloride, approximately 10 mOsm for 300 mg/L NaCl or 400 mg/L of KCl

Sodium Bicarbonate Amount

   Powdered Media Product No.      NaHCO3 7.5% Solution (mL/L)      NaHCO3 Powder (g/L)
Ames' Medium A1420 25.7 1.932
BME EBSS B9638 29.3 2.2
DME/F-12 w/o phenol red D2906 16 1.2
DME/F-12 D9785 16 1.2
DME/F-12 D8900 16 1.2
DME/F-12 D0547 16 1.2
DME HEPES D1152 49.3 3.7
DME w/o phenol red D2902 49.3 3.7
DME Deficient D5030 49.3 3.7
DME D5523 49.3 3.7
DME D5648 49.3 3.7
DME D7777 49.3 3.7
F-12 (Coon's Modification) F6636 35.7 2.676
Glasgow MEM G6148 36.7 2.75
HBSS w/o phenol red H1387 4.7 0.35
HBSS H2387 4.7 0.35
HBSS w/o phenol red H4891 4.7 0.35
HBSS H6136 4.7 0.35
IMDM I7633 40.3 3.024
KREBS-HENSELEIT K3753 28 2.1
KREBS-RINGER K4002 16.8 1.26
McCOY'S 5A M4892 29.3 2.2
MCDB-131 M8537 15.7 1.176
MCDB-153 M7403 15.7 1.176
M199 EBSS HEPES M2520 29.3 2.2
M199 EBSS w/o phenol red M3769 29.3 2.2
M199 EBSS M5017 29.3 2.2
M199 HBSS M0393 4.7 0.35
MEM EBSS M0268 29.3 2.2
MEM JOKLIK M0518 26.7 2
MEM EBSS NEAA M0643 29.3 2.2
MEM Alpha M0644 29.3 2.2
MEM Alpha M0894 29.3 2.2
MEM HBSS NEAA M1018 4.7 0.35
MEM EBSS HEPES M2645 29.3 2.2
MEM EBSS NEAA w/o phenol red M3024 29.3 2.2
MEM HBSS M4642 4.7 0.35
MEM EBSS Automod M0769 29.3 2.2
NCTC 135 N3262 29.3 2.2
Nutrient Mixture F-10 N6635 16 1.2
Nutrient Mixture F-12 N6760 15.7 1.176
Nutrient Mixture F-12K N3520 33.3 2.5
RPMI-1640 Deficient R1383 26.7 2
RPMI-1640 HEPES R4130 26.7 2
RPMI-1640 R6504 26.7 2
RPMI-1640 Hybrimax R8005 26.7 2
RPMI-1640 Automod R7755 26.7 2
RPMI-1640 w/o phenol red R8755 26.7 2
Tyrode's Salts T2145 13.3 1
Waymouth MB752/1 W1625 29.9 2.24
Williams' Medium E W4125 29.3 2.2

Materials

     
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